The Revised Diploid Genome Sequence of an Individual Human: An Optimized Assembly Workflow for Scaling of near Telomere-to-Telomere Assemblies

The first draft diploid genome assembly (HuRef) of an individual released in 2007 was a milestone in genomics. Here, we report HuRef2.0, a revision of HuRef assembled using a scalable two-step workflow employing only Oxford Nanopore Technologies (ONT) Simplex reads and the hifiasm assembler. Results are close in continuity to the recent telomere-to-telomere (T2T) assemblies, but were assembled from standard DNA samples without using multiple sequencing and mapping technologies, including ultra-long-reads and/or proximity-ligation. Three ONT flowcells ([~]103x coverage) from fresh blood DNA produced an assembly comprising 26 contigs, with gapless assembly of 23 chromosomes. Two gaps on chromo